Glycinebetaine (GB) is an important organic osmolyte that accumulates in many plant species in response to abiotic stresses including heavy metals. The objective of this study was to investigate whether exogenous GB would ameliorate the adverse effect of cadmium (Cd) stress on perennial ryegrass (Lolium perenne). Fifty-three days old seedlings were exposed to hydroponic culture for 7 days with six treatments: T1 (control), T2 (0 mM Cd + 20 mM GB), T3 (0 mM Cd + 50 mM GB), T4 (0.5 mM Cd + 0 mM GB), T5 (0.5 mM Cd + 20 mM GB), T6 (0.5 mM Cd + 50 mM GB). Cd stress resulted in a remarkable decrease in turf quality, vertical shoot growth rate (VSGR), normalized relative transpiration (NRT) and Chlorophyll (Chl) content; with significant increases in electric conductivity (EL), malondialdehyde (MDA) content, superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) activity, oxalic and tartaric acid content. Exogenous application of GB decreased EL and MDA content in Cd stressed plants, and increased turf quality, VSGR, NRT, Chl content, SOD, CAT, POD activity, oxalic, tartaric acid content, and the gene expression level of SOD and POD when compared with Cd stressed without GB. Perennial ryegrass with 20 mM GB application suppressed the Cd accumulation in both shoots and roots. A lower translocation factor of Cd was found in GB treated plants than non-GB treated plants, and the lowest translocation factor was observed in the 20 mM GB application. These results suggested that GB could alleviate the detrimental effect of Cd on perennial ryegrass and the amelioration was mainly related to the elevation in SOD, CAT, and POD at enzyme and gene expression levels, which reduced Cd content in shoots and improved cell membrane stability by reducing oxidation of membrane lipids. These findings lead us to conclude that application of GB with 20 mM is the best strategy to ameliorate the detrimental impacts of Cd stress on perennial ryegrass.